The impact of various carbon nanomaterials on the morphological, behavioural, and biochemical parameters of rainbow trout in the early life stages.

With the increasing production and the number of potential applications of carbon nanomaterials, mainly from the graphene family, their release into the natural environment, especially to aquatic ecosystems, is inevitable. The aim of the study was to determine the effects of various carbon nanomaterials (graphene nanoflakes (GNF), graphene oxide (GO), reduced graphene oxide (RGO) and silicon carbide nanofibers (NFSiC) in the concentration of 4 mg L-1 on the early life stages of the rainbow trout Oncorhynchus mykiss. The survival rates of O. mykiss were not affected after 36 days of exposure to studied materials, except for RGO, which caused significant mortality of both embryos and larvae compared to the control conditions. Larvae exposed to GO and NFSiC were characterized by a smaller standard body length at hatch, whereas at the end of the experiment, the growth of fish exposed to all materials was accelerated, especially in GO and RGO treatment, in which higher body weight and length were accompanied by lower volume of the yolk sac. Neither the markers of the oxidative damage nor the antioxidant enzymes activities were significantly affected in embryos, newly hatched larvae and larvae after 26-day exposure to studied carbon nanomaterials. Also, no neurotoxic effect expressed by the activity of the whole-body acetylcholinesterase was observed. Nevertheless, the significant increase in the velocity and the overall activity of larvae exposed to GNF (not investigated after exposure to other materials) must be highlighted. The most pronounced effect of RGO might be connected with its large particle size, sharp edges, and the presence of TiO2 nanoparticles. The results indicate for the first time that various carbon nanomaterials potentially released into aquatic ecosystems may have serious developmental implications for the early life stages of salmonid fish.

Do magnetic fields related to submarine power cables affect the functioning of a common bivalve?

The aim of the study was to determine the effect of static magnetic field (SMF) and electromagnetic field (EMF), of values usually recorded near submarine cables, on the bioenergetics, oxidative stress, and neurotoxicity in the cockle Cerastoderma glaucum. Bivalves maintained a positive energy balance, but the filtration rate and energy available for individual production were significantly lower in SMF-exposed animals compared to the control treatment. No changes in the respiration were noted but ammonia excretion rate was significantly lower after exposure to EMF. Changes in the activities of antioxidant enzymes and the lipid peroxidation were not observed however, exposure to both fields resulted in increased protein carbonylation. After exposure to EMF a significant inhibition of acetylcholinesterase activity was observed. As the present study for the first time revealed the oxidative damage and neurotoxicity in marine invertebrate after exposure to artificial magnetic fields, the need for further research is highlighted.

Microplastics cause neurotoxicity and decline of enzymatic activities in important bioturbator Hediste diversicolor.

Microplastics (MPs) tend to accumulate in marine sediments thus benthic fauna is particularly vulnerable to microplastic pollution. Hediste diversicolor is a widespread species in coastal marine sediments. It plays key ecological functions mostly related to bioturbation process which means sediment reworking due to the worm burrowing activity and building a network of galleries. Herein, we show that commercial plastic microspheres of two sizes (63-75 and 300-355 µm) have the potential to cause neurotoxicity in H. diversicolor. The whole-body acetylcholinesterase (AChE) activity - a common indicator of neurotoxic effect - was on average 60% lower in polychaetes exposed for 28 days to MPs served at environmentally relevant concentrations (0.08% sediment d. wt.), than in unexposed ones. Significantly reduced activities of antioxidant enzymes (SOD, CAT, GST) indicated suppression of the cellular antioxidative system in worms exposed to MPs. No changes were, however, observed in tGSH, lipid or protein oxidation measures (CBO, MDA), and in the energetic value of these polychaetes. The response was generally similar with no regard to MPs size. Only very few microspheres were found in polychaetes exposed to MPs spiked sediment. The potential role of MPs-associated pollutants as a factor responsible for observed biochemical effects, is discussed.

Plastic in digestive tracts and gills of cod and herring from the Baltic Sea.

Plastic litter is widespread on our planet and is recognized as a contaminant of high concern. Plastic ingestion and retention in gills were studied in two key Baltic fish species: herring (Clupea harengus L.) and cod (Gadus morhua L.). In total, 183 fish from the southern Baltic Sea were analysed. Plastic litter was found in digestive tracts of 12.7 and 14.8% of herrings and cods, respectively. In addition, gills were shown to constitute an important transfer route of plastic to cod (9.9% of cods) but not to herring thus likely reflecting species-specific differences in fish lifestyles and/or water filtering capacity. No more than one plastic item per individual was found except for three fish with two items in their stomachs. Dominant microplastics (<5 mm) (MPs) (88.6% of all items) composed of irregular fragments, fibres and foils (61.3%, 25.8% and 12.9% of all MPs, respectively) were followed by meso- (5-25 mm) and macro-plastics (>25 mm) (5.7% each of all items). Plastic ingestion and retention in gills did not affect fish body condition assessed by Fulton's K index, although herrings that ingested plastic particles beyond the microplastic size range (i.e. >5 mm) were characterized by the lowest condition indices. All plastic items recovered from fish exhibited clear signs of weathering and surface extended due to their roughness and fragmentation which may enhance both sorption properties for chemical compounds and/or their leaching. Nevertheless, chemical contaminant levels in herring muscles were not different than in a plastic-free group. The vast majority of fibres found in fish was identified as air-borne or procedural contamination and were excluded from further analyses. In addition, the vessel's paint dust found in fish stressed the need for strict quality assurance also during fish sampling.

Effects of different types of primary microplastics on early life stages of rainbow trout (Oncorhynchus mykiss).

Plastic pollution is recognized as serious threat to aquatic organisms. The aim of this research was to determine the effect of environmentally realistic concentrations of various microplastics (MPs) on survival, growth, development and induction of endocrine, geno- and cytotoxic responses in the early life stages of rainbow trout Oncorhynchus mykiss. Fish were exposed for 69-days, from embryos at eyed-stage to mobile yolk-sac larvae, to pre-production pellets (3000 µm; polystyrene - PS and polyethylene terephthalate - PET). Additionally, since salmonid larvae are particularly exposed to light polymers after swimming up from the bottom, fish were also treated with PE microspheres (150-180 µm; polyethylene - PE) for both long (69-days, from embryos at eyed-stage) and short period (29 days, from larvae 3 weeks after hatching) to test the development stage-related effect on the growth parameters and fitness. Hatching success, rate and the survival of larvae did not differ among treatments. Although some alterations were found in the length gain after the long-term exposure and in the yolk-sac exhaustion rate in all PE treatments, the final size of larvae did not differ from the respective controls. PE-treated larvae have shown elevated corticosterone concentrations being significantly higher in fish exposed from the embryo stage. It was indicated for the first time that mobile yolk-sac larvae ingested MPs (up to 24% of larvae contained microspheres). No changes were recorded in cytotoxicity endpoints in any of the treatments, but exposure to PS pellets resulted in significantly higher frequencies of genotoxicity endpoints compared to the control treatment. This effect and aforementioned alterations in PE-treated larvae might result from the exposure to toxic MPs leaches. The fact that selected PAHs' levels reached the highest values in PS pellets and PE microspheres must be underlined.

Response of sediment-dwelling bivalves to microplastics and its potential implications for benthic processes.

Sediments are considered an ultimate sink for microplastic (MPs) but knowledge on MPs impacts on the benthic organisms is still limited. A microcosm experiment was performed to assess the effect of MPs on two sediment-dwelling bivalve species that differ in terms of their life¬styles and feeding behaviors (the cockle Cerastoderma glaucum and the Baltic clam Limecola balthica). Fitness of both bivalves, their vertical distribution in the sediment and activity at the sediment-water interface were studied following addition of PE microspheres of three different size fractions (63-75, 150-180 and 250-300 µm) in two concentrations (0.1 and 0.5% sediment dwt) to the sediment surface. Bivalve survival was generally high and did not vary significantly among treatments. Both bivalve species captured MPs but no significant effect on their body conditions and energy reserves was observed. However, some behavioural alterations were found. The near-surface-dwelling C. glaucum emerged from the sediment less often and in lower numbers suggesting lower activity in sediments amended with large and medium MPs served in higher concentration. Also vertical distribution of deeper-dwelling L. balthica differed among treatments. The clam penetrated deeper sediment layers in microcosms treated with MPs than in the control (on average 62% of the total number of bivalves found in the sediment layer 2-4 cm vs. 65% in the upper 2 cm) which may suggest an avoidance behaviour or escape reaction in response to MPs addition to the topmost sediment. Total sediment community oxygen consumption was significantly higher in MPs-amended sediments than in the control, with the highest rates in treatment with the largest microspheres. However, oxygen uptake increase seemed to be unrelated to addition of MPs per se, suggesting rather an interaction between MPs and bivalves. The need for future studies on MPs effects on benthic community structure and functioning is highlighted.

Do the graphene nanoflakes pose a potential threat to the polychaete Hediste diversicolor?

Graphene is a promising material with a wide range of future applications that could potentially lead to its transfer from numerous water and terrestrial sources to the sea, thus fate and effects of graphene in the marine ecosystem deserve attention. Within this work, the impact of the short- and long-term exposure (36 h and 24 days) of the marine benthic polychaete Hediste diversicolor to various concentrations (36 h: 0.4, 4, 40 and 400 mg L-1; 24 days: 4 and 40 mg L-1) of the pristine graphene multilayer nanoflakes (of thickness 8-12 nm) was investigated. Experiments revealed a limited toxic effect of graphene on H. diversicolor. Although the polychaetes ingested graphene, no impact on their total energy content was found. The toxic effect expressed by significant elevation of catalase activity indicating activation of defence mechanisms was recorded but only at the early stage of exposure. Activities of other antioxidant and cellular damage biomarkers (SOD, GST, GSH, MDA, CBO) remained unaffected. Moreover, no neurotoxic effect expressed by inhibition of acetylcholinesterase (AChE) activity was observed. Substantial inter-individual variability in the activities of some biomarkers at the end of the long-term experiment was found. Polychaetes were buried deeper in the sediment with graphene than in the controls indicating an escape reaction and avoidance behaviour. The latter may lead to the transfer of graphene from the sediment surface to deeper sediment layers with unknown consequences for the benthic ecosystem.

Effects of chronic exposure to microplastics of different polymer types on early life stages of sea trout Salmo trutta.

The aim of the present study was to determine the effect of a long-term (113 days) exposure to microplastics on the development and induction of endocrine, geno- and cytotoxic responses in early life stages of sea trout Salmo trutta. Microplastic particles (3000 µm) of three most commonly mass-produced polymers (polystyrene - PS, polyethylene terephthalate - PET and polyethylene - PE) were applied in environmentally realistic concentrations (0.1% of sediment dry weight) in a laboratory experiment imitating the natural environment, typical for sea trout spawning grounds. The exposure of the sea trout, from fertilized eggs to mobile yolk-sac larvae, to microplastics did not affect the hatching success (the survival of embryos), hatching rate and the incubation period. Microplastics of any tested polymer type also had no adverse effect on the larvae survival, growth rate and the rate of yolk sack absorption. Similarly, no changes in frequencies of detected cytotoxicity endpoints compared to the control group were recorded. Exposure to polymer particles induced however the formation of genotoxicity endpoints (nuclear buds, micronuclei and blebbed nuclei cells). The level of total genotoxicity (ΣGentox) in fish larvae erythrocytes increased significantly in the following sequence: PS > PET > PE. No significant changes in the whole body corticosterone, dehydrocorticosterone and cortisone concentrations due to exposure to microplastics were recorded, while cortisol was detected in larvae exposed to PS. Our results show that long-term, non-ingestion related exposure to microplastics does not affect development of S. trutta early life stages but may lead to genotoxic responses. PS seems to be the most hazardous among all polymers studied. This is the first study demonstrating non-ingestion related toxicity of microplastics to the early life stages of fish.

Microplastics on sandy beaches of the southern Baltic Sea.

Microplastic occurrence and composition were investigated along the Polish coast (southern Baltic Sea) on 12 beaches differing in terms of intensity of their touristic exploitation, urbanisation and sediment characteristics. Their mean concentrations varied between 76 and 295 items per kg dry sediment. Fibres and plastic fragments were the dominant microplastic types. Overall, no relationship was found between their concentrations and sediment characteristics. Fine sediments were not identified as microplastic pollution traps. The highest microplastic concentrations were recorded at some urban beaches indicating that population density and the level of coastal infrastructure development are important factors affecting microplastic pollution level on beaches. On the other hand, microplastic concentrations in national parks did not differ substantially from the other beaches. Our results suggest that sediment accumulation processes may exceed microplastic accumulation, and overcome the effect of tourism and/or urbanisation, highlighting the role of the beach hydrodynamic status in structuring beach microplastic pollution.

Effect of low frequency electromagnetic field on the behavior and bioenergetics of the polychaete Hediste diversicolor.

The aim of the present study was to determine the effect of an electromagnetic field (EMF) of value typically recorded in the vicinity of submarine cables (50 Hz, 1 mT) on the behavior and bioenergetics of the polychaete Hediste diversicolor. No avoidance or attraction behavior to EMF was shown, but the burrowing activity of H. diversicolor was enhanced in EMF treatment, indicating a stimulating effect on bioturbation potential. The polychaete maintained a positive energy balance and high amount (85% of assimilated energy) of energy available for individual production (scope for growth) after exposure to EMF. Food consumption and respiration rates were not affected but ammonia excretion rate was significantly reduced in EMF-exposed animals compared to the control conditions (geomagnetic field). The mechanisms behind this effect remain, however, unclear. This is the first study demonstrating the effects of environmentally realistic EMF value on the behavior and physiology of marine invertebrates, thus there is a need for more research.

Are magnetic and electromagnetic fields of anthropogenic origin potential threats to early life stages of fish?

The number of underwater cables transferring electric current in sea and freshwater environments is constantly increasing. As a result, the risk of negative effects of magnetic fields generated in the vicinity of those cables on fish eggs and larvae is also growing. This is especially the case for species that settle on the bottom for certain periods of time during early development. To study those effects, eggs and larvae of rainbow trout, Oncorhynchus mykiss, were subjected under experimental conditions to a static magnetic field (MF) of 10 m T and a 50 Hz electromagnetic field (EMF) of 1 m T for a period of 36 days (i.e., from eyed egg stage to approximately 26 days post hatching, dph). Neither MF nor EMF had significant effect on embryonic or larval mortality, hatching time, larval growth, or the time of larvae swim-up from the bottom. However, both MF and EMF enhanced the yolk-sac absorption rate. Although it was not related directly to magnetic field effect, it was also shown that larvae with absorbed yolk-sacs by the time of swim-up were less efficient in taking advantage of available food at first feeding (i.e., obtained smaller weight at age). That indicates the importance of processes affecting yolk-sac absorption rate.

Genotoxic and cytotoxic effects of 50 Hz 1 mT electromagnetic field on larval rainbow trout (Oncorhynchus mykiss), Baltic clam (Limecola balthica) and common ragworm (Hediste diversicolor).

The aim of this research was to assess genotoxicity and cytotoxicity responses in aquatic animals exposed to 50 Hz 1 m T electromagnetic field (EMF). Rainbow trout (Oncorhynchus mykiss) at early stages of development were exposed to EMF for 40 days, whereas marine benthic invertebrates - the common ragworm Hediste diversicolor and the Baltic clam Limecola balthica - for 12 days. To define genotoxicity and cytotoxicity responses in selected animals, assays of nuclear abnormalities in peripheral blood erythrocytes of O. mykiss, coelomocytes of H. diversicolor and gill cells of L. balthica were performed. Induction of formation of micronuclei (MN), nuclear buds (NB), nuclear buds on filament cells (NBf) and cells with blebbed nuclei (BL) were assessed as genotoxicity endpoints, and 8-shaped nuclei, fragmented (Fr), apoptotic (Ap) and binucleated (BN) cells as cytotoxicity endpoints. Exposure to EMF affected all studied species but with varying degrees. The strongest responses to EMF treatment were elicited in L. balthica, in which six out of the total eight analyzed geno- and cytotoxicity endpoints were significantly elevated. Significantly induced frequencies of MN were detected in O. mykiss and H. diversicolor cells, NBf and BL only in gill cells of L. balthica, and NB in analyzed tissues of all the test species. As cytotoxicity endpoints, a significant elevation in frequencies of cells with 8-shaped nuclei was found in O. mykiss and L. balthica, while Ap and BN was observed only in L. balthica. EMF exposure did not induce any significant cytotoxic activity in H. diversicolor coelomocytes. The present study is the first to reveal the genotoxic and cytotoxic activity of 1 m T EMF in aquatic animals, and, consequently, the first one to report the adverse effect of this factor on common marine invertebrates and early life stages of fish.

Marine litter on the seafloor of the southern Baltic.

Marine litter occurrence and composition were investigated during routine bottom trawl fish surveys type BITS performed in the Polish Maritime Areas (the southern Baltic Sea). Sampling covered a distance of 325km and an area of 16km2 at a depth range of 19-110m. Litter densities varying between 0 items/ha (34% of tows) and 2.23items/ha with a mean of 0.20items/ha (SD=0.30) are at the bottom range of densities reported from other shelf habitats worldwide at similar water depths. The majority of the items (40%) were found at a depth range of 51-60m. Overall, plastic was the most common litter type (67% of all items) found in all tows with litter. The results of this study indicate that despite the Baltic being a semi-enclosed basin, with a densely populated coastline and extensive shipping, marine litter pollution of the southern Baltic seafloor is low compared to other coastal areas.

Species-specific effect of macrobenthic assemblages on meiobenthos and nematode community structure in shallow sandy sediments.

Three functionally different macrofaunal species (the filter- and/or surface deposit-feeding polychaete Hediste diversicolor, and the suspension-feeding bivalves Mya arenaria and Cerastoderma glaucum) were introduced as single- and two-species treatments into microcosms containing sandy sediment with a natural meiofaunal community. H. diversicolor is a burrowing species building a system of galleries, C. glaucum lives actively near the sediment surface acting as a biodiffuser and M. arenaria buries deeply and leads a sessile lifestyle. It is shown that H. diversicolor extended the vertical distribution of meiofauna into deeper sediment layers compared to the control and non-Hediste treatments. The response of the nematode community varied significantly among treatments and was dependant on the macrobenthic species composition but not on the species number. Nematode assemblages in all treatments with the polychaete, both in monoculture and with either bivalve, differed significantly from those recorded in other treatments and were more similar than replicates within any other single treatment. H. diversicolor also appeared to have stimulated nematode species diversity. The present study demonstrated that the impact of macrobenthic assemblages on meiofauna is not a simple summation of individual species effects but is species specific.

The oxygen fluxes of sandy littoral areas: quantifying primary and secondary producers in the Baltic Sea.

The study aimed to estimate the total oxygen flow through the shallow littoral of the Gulf of Gdansk (southern Baltic Sea). Daily primary production and daily oxygen consumption by meso- and macroplankton and meio- and macrobenthos were measured, as well as the remaining (abiotic/microbial) oxygen consumption of the water column and bottom sediment. The daily gross primary production was 5.0 l of oxygen m(-2), oxygen consumption was 9.2 l m(-2), of which 16% were used by the water column and 84% by the bottom sediment. Abiotic oxygen consumption in water column contributed to less than 5% of the total oxygen consumption, and in the bottom sediment to 67%. Mesoplankton in the water column and meiobenthos in the bottom sediments were the major oxygen consumers in the littoral (67% and 26%, respectively).